CN108918797A - The reagent method and reagent chip of a kind of blood platelet method for separating and concentrating, Platelet drug - Google Patents
The reagent method and reagent chip of a kind of blood platelet method for separating and concentrating, Platelet drug Download PDFInfo
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- CN108918797A CN108918797A CN201810504137.9A CN201810504137A CN108918797A CN 108918797 A CN108918797 A CN 108918797A CN 201810504137 A CN201810504137 A CN 201810504137A CN 108918797 A CN108918797 A CN 108918797A
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Abstract
The reagent method and reagent chip of a kind of blood platelet method for separating and concentrating, Platelet drug, are related to platelets analysis field.Blood platelet method for separating and concentrating is to flow blood sample in the duct, applies dielectrophoresis force；Inducer is added in blood platelet；Flow mixture in the duct, apply dielectrophoresis force, the quantity of detection of aggregation blood platelet and blood platelet, the blood platelet method for separating and concentrating can quickly, efficiently separate out blood platelet in blood sample.The reagent method of Platelet drug is that Platelet drug is added in the mixture；Mixture after making drug effect flows in the duct, applies dielectrophoresis force, the quantity of detection of aggregation blood platelet and blood platelet, and the reagent method of the Platelet drug can quickly detect the Assembling Behavior variation after blood platelet and drug effect.The structure of the reagent chip of Platelet drug is simple, at low cost, can quickly carry out the reagent of Platelet drug.
Description
Technical field
The present invention relates to platelets analysis fields, and in particular to a kind of blood platelet method for separating and concentrating, Platelet
The reagent method and reagent chip of drug.
Background technique
Blood platelet participates in many physiology, pathologic process in body as peripheral blood important component.In blood coagulation physiology course
In, in blood, a small amount of blood platelet will be attached on extracellular matrix protein first for extracellular matrix protein exposure at injury of blood vessel
Surface, subsequent blood platelet are activated, by the coagulation factors such as the adenosine diphosphate (ADP) (ADP) of autocrine and thromboxane A2 (TXA2) from
Other blood platelets are raised in blood, combine eventually by II b/ of GP, the III a receptor and fibrinogen of blood platelet and aggregation occurs instead
It answers.The inhibition of platelet adhesion reaction aggregation capability will lead to bleeding risk, and hyperfunction will increase atherosclerosis, coronary artery
The thrombus diseases risk such as disease, myocardial infarction, stroke.Therefore, blood platelet physiological function, especially its aggregation are measured, not only
Help to diagnose hemorrhagic disease caused by certain congenital and acquired platelet defect, and in thrombus disease morbidity machine
It is significant in the research such as system, clinical diagnosis, antithrombotic therapy, monitoring anti-platelet drug concentration.
Currently, platelet aggregation has become clinical one of basic detection project, existing detection technique and system packet
Include photoelectric turbidimetry, whole blood electric-resistivity method, PFA-100 platelet function instrument, PL-11 whole bliid platelet analyzer, Verifynow system
And thrombelastogram instrument etc..Wherein whole blood dynamic analysis platelet aggregation can be used directly simultaneously in PL-11 whole bliid platelet analyzer
Can various ingredients (including blood platelet and red blood cell) to blood cell carry out quantitative analysis.
For the medication effect of effective, the accurate Platelet drug for obtaining individual patient, need to detect individual trouble
Assembling Behavior variation after blood platelet and drug effect in person's blood.And above-mentioned existing detection technique and system are all very multiple
It is miscellaneous, it is at high cost, it is only suitable for professional detection department, it is clear that be not suitable for quickly carrying out reagent process.In addition, being needed for quick reagent
It first to be come out to by the blood platelet quick separating in individual patient blood, but existing blood platelet separation method and equipment are same
In the presence of complicated, at high cost, the defect of destructible blood platelet.
Therefore, it is necessary to it is a kind of can quick separating go out the blood platelet in blood, and then the method and apparatus for carrying out reagent.
Summary of the invention
The purpose of the present invention is to provide a kind of blood platelet method for separating and concentrating, the method can quickly, efficiently separate out
Blood platelet in blood sample.
Another object of the present invention is to provide a kind of reagent methods of Platelet drug, and it is small to quickly detect blood
Assembling Behavior after plate and drug effect changes.
Another object of the present invention is to provide a kind of reagent chip of Platelet drug, structure is simple, cost
It is low, it can quickly carry out the reagent of Platelet drug.
The present invention solves its technical problem and adopts the following technical solutions to realize.
The present invention proposes a kind of blood platelet method for separating and concentrating comprising following steps：
It flows blood sample in the duct, dielectrophoresis force is applied to the blood sample sample in pipeline, is made in blood sample
Blood platelet and other particles separate, detect the quantity of blood platelet；
Inducer is added in blood platelet, assembles blood platelet sufficiently, obtains the mixture of aggregation blood platelet and blood platelet；
Make to assemble blood platelet and the mixture of blood platelet flows in the duct, dielectrophoresis is applied to the mixture in pipeline
Power separates aggregation blood platelet and blood platelet in mixture, the quantity of detection of aggregation blood platelet and the quantity of blood platelet.
Further, in a preferred embodiment of the present invention, the method for detection blood platelet/aggregation blood platelet quantity is：It adopts
With Electrode with Electrochemical Impedance Spectroscopy, blood platelet/aggregation blood platelet electrochemical impedance and frequency are detected, according to blood platelet/aggregation blood platelet
Electrochemical impedance and frequency and blood platelet/aggregation platelet counts between corresponding relationship, obtain blood platelet/aggregation blood platelet
Quantity.
Further, in a preferred embodiment of the present invention, the frequency of the dielectrophoresis force of application is 500KHz-3MHz.
Further, in a preferred embodiment of the present invention, inducer include collagen, adenosine diphosphate (ADP), adrenaline,
One of arachidonic acid, ristomycin.
Further, in a preferred embodiment of the present invention, flow blood sample or mixture in pipeline by air pump.
A kind of reagent method of Platelet drug comprising following steps：
Using above-mentioned blood platelet method for separating and concentrating, the quantity of aggregation blood platelet and the quantity of blood platelet are obtained；
Platelet drug, the aggregation after obtaining drug effect are added in the mixture of aggregation blood platelet and blood platelet
The mixture of blood platelet and blood platelet；
Mixture after making drug effect flows in the duct, applies dielectrophoresis force to the mixture in pipeline, makes medicine
Object effect after mixture in aggregation blood platelet and blood platelet separate, detect drug effect after assemble blood platelet quantity with
The quantity of blood platelet.
Further, in a preferred embodiment of the present invention, Platelet drug is divided into four classes, respectively inhibition blood platelet
Arachidonic acid metabolic medicine, the medicine for increasing blood platelet cyclic nucleotide content, specificity inhibit medicine, the blood of ADP activated blood platelet
Platelet fibrinogen deceptor antagonists.
A kind of reagent chip of the Platelet drug of the reagent method based on above-mentioned Platelet drug, packet
Main body is included, three branches is equipped in main body and a pipeline, one end of three branches is connected to the same end of pipeline respectively, pipeline
Be connected with one end of three articles of branches be followed successively by the other end for making that the blood platelet in blood sample and other particles separate
One separation and concentration area, the first quantity detection zone for detecting platelet counts, the aggregation for assembling blood platelet sufficiently lure
Lead area, the second separation and concentration area for separating aggregation blood platelet and blood platelet in mixture, for detection of aggregation blood it is small
It is second quantity detection zone of the quantity of the quantity and blood platelet of plate, reagent area, poly- in the mixture after drug effect for making
The third separation and concentration area that is separated with blood platelet of collection blood platelet and for detect the quantity for assembling blood platelet after drug effect and
The quantity third quantity detection zone of blood platelet.
Further, in a preferred embodiment of the present invention, offered in main body respectively with three branches far from pipeline one
Hold injection port, the first reagent wells, the second reagent wells of connection, the inducer adding mouth being connected to aggregation inducing area, with reagent area
The drug adding mouth of connection, the outlet being connected to pipeline far from one end of three branches.
Further, in a preferred embodiment of the present invention, pipeline successively include along its length by the first separation and concentration area,
First quantity detection zone and aggregation inducing district's groups are at the first segment pipe, the first connecting pipe, by the second separation and concentration area, the second number
Measure detection zone and reagent district's groups at the second segment pipe, the second connecting pipe detects by third separation and concentration area and third quantity
District's groups at third segment pipe, the valve of control pipeline opening and closing is separately provided in the first connecting pipe and the second connecting pipe
Door.
The blood platelet method for separating and concentrating of the embodiment of the present invention, the reagent method of Platelet drug and reagent chip
Beneficial effect is：The blood platelet method for separating and concentrating of the embodiment of the present invention is to flow blood sample in the duct, in pipeline
Blood sample sample apply dielectrophoresis force, separate blood platelet and other particles in blood sample, detect the quantity of blood platelet；
Inducer is added in blood platelet to stay for some time, assembles blood platelet sufficiently, obtains the mixed of aggregation blood platelet and blood platelet
Close object；Make to assemble blood platelet and the mixture of blood platelet flow in the duct, dielectrophoresis force is applied to the mixture in pipeline,
Separate aggregation blood platelet and blood platelet in mixture, the quantity of detection of aggregation blood platelet and the quantity of blood platelet, the blood is small
Plate method for separating and concentrating can quickly, efficiently separate out blood platelet in blood sample.The Platelet of the embodiment of the present invention
The reagent method of drug is added in the mixture of aggregation blood platelet and blood platelet after the completion of blood platelet method for separating and concentrating
Platelet drug, the mixture of aggregation blood platelet and blood platelet after obtaining drug effect；Mixing after making drug effect
Object flows in the duct, applies dielectrophoresis force to the mixture in pipeline, the aggregation blood in the mixture after making drug effect
Platelet is separated with blood platelet, the quantity of the aggregation blood platelet after detecting drug effect and the quantity of blood platelet, the Platelet
The reagent method of drug can quickly detect the Assembling Behavior variation after blood platelet and drug effect.The blood of the embodiment of the present invention is small
The structure of the reagent chip of plate drugs with function is simple, at low cost, can quickly carry out the reagent of Platelet drug.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is seperated schematic diagram of the blood sample under dielectrophoresis force effect in the embodiment of the present invention；
Fig. 2 is a kind of structural schematic diagram of the reagent chip of Platelet drug provided in an embodiment of the present invention；
Fig. 3 is the structural schematic diagram of Tu2Zhong branch and pipe section.
Icon：100- reagent chip；110- main body；120- branch；130- pipeline；141- injection port；The first reagent of 142-
Hole；The second reagent wells of 143-；144- inducer adding mouth；145- drug adding mouth；146- outlet；The first segment pipe of 150-；
The first separation and concentration of 151- area；152- the first quantity detection zone；153- aggregation inducing area；The first connecting pipe of 160-；161- valve
Door；The second segment pipe of 170-；The second separation and concentration of 171- area；172- the second quantity detection zone；173- reagent area；180- second
Connecting pipe；190- third segment pipe；191- third separation and concentration area；192- third quantity detection zone.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Below to the reagent method and reagent of the blood platelet method for separating and concentrating of the embodiment of the present invention, Platelet drug
Chip is specifically described.
The embodiment of the present invention provides a kind of blood platelet method for separating and concentrating comprising following steps：
S1, it flows blood sample in the duct, dielectrophoresis force, the dielectric of application is applied to the blood sample sample in pipeline
The frequency of electrophoretic force is generally 500KHz-3MHz, separates blood platelet and other particles in blood sample.Usually in 0.5-
Sheath fluid is added in the blood sample of 5ml, and flows blood sample in the pipeline of diameter 15-50um, flowing velocity is according to reality
It is under normal conditions 10-200ul/min depending on the situation of border.
It is shown in Figure 1, according to laminar flow theory, the liquid flow that is flowed in microchannel and tube wall, process and process it
Between viscous force lead at tube wall that process flow velocity is minimum, flow velocity is maximum at pipe axis, speed and pipe axis bigger closer to pipe axle speed
The distance at place is at parabolic distribution.When flow field is to illustrate direction flowing, electrod-array is placed in runner bottom, forms non-homogeneous electricity
, by selecting suitable frequency to make blood cell and blood platelet by dielectrophoresis force, due to dielectrophoresis force and particle
Radius is directly proportional, and the particle of different scale will bear different dielectrophoresis forces, their equilbrium positions in vertical direction
Also different, therefore variable grain is respectively at the different layers of parabolic velocity profile, therefore has different certain movement speeds.
By selecting suitable frequency that blood platelet can be made to separate with other particles, flows, collected by timesharing at different rates
Blood platelet or other particles can be respectively obtained.In general, it can be acted on by inhomogeneous field so that blood platelet speed
Faster, to first be collected into blood platelet.
S2, the quantity for detecting blood platelet, specific method are：Using Electrode with Electrochemical Impedance Spectroscopy, the electrochemistry resistance of blood platelet is detected
Anti- and frequency obtains blood platelet according to the corresponding relationship between the electrochemical impedance and frequency and platelet counts of blood platelet
Quantity.
According to Electrode with Electrochemical Impedance Spectroscopy principle, the component part of cell includes cell membrane, cytoplasm, nucleus, different groups
There are different dielectric properties at part, such as, cell membrane be it is nonconducting, cytoplasm has good conduction
Performance uses impedance spectrum, the cell and particle of measurement frequency response by creating equivalent-circuit model；Pass through different cells
Property, such as thickness and the cytoplasmic conductivity of cell membrane, cell is distinguished；For the cell or grain of different number
Son, corresponding electrochemical impedance and frequency are accordingly also different, and corresponding corresponding relationship is presented.
S3, in blood platelet be added inducer make platelet aggregation, and stay for some time (0-300s) fill blood platelet
Divide aggregation, obtains the mixture of aggregation blood platelet and the blood platelet that do not assemble.The inducer that the present embodiment uses includes collagen egg
White (COLL), adenosine diphosphate (ADP) (ADP), adrenaline (EPN), arachidonic acid (ACA), in ristomycin (RIST)
It is a kind of.
S4, flow the mixture of aggregation blood platelet and the blood platelet that do not assemble in the duct, to the mixture in pipeline
Apply dielectrophoresis force, the frequency of the dielectrophoresis force of application is generally 500KHz-3MHz, keeps the aggregation blood in mixture small
Plate is separated with the blood platelet that do not assemble.
The quantity of S5, the quantity of detection of aggregation blood platelet and the blood platelet that do not assemble, specific method are：It is hindered using electrochemistry
Anti- method detects blood platelet/aggregation blood platelet electrochemical impedance and frequency, is hindered according to blood platelet/aggregation blood platelet electrochemistry
The anti-and corresponding relationship between frequency and blood platelet/aggregation platelet counts, obtains blood platelet/aggregation blood platelet quantity.
The blood platelet method for separating and concentrating of the present embodiment is carried out in a pipeline, and pipeline successively wraps along its length
5 regions are included, and flow blood sample or mixture in pipeline by air pump, i.e., from a next area of field flow orientation
Domain.5 regions are respectively：The first separation and concentration area for separating the blood platelet in blood sample with other particles is used for
Detect the first quantity detection zone of platelet counts, the aggregation inducing area for assembling blood platelet sufficiently, for making mixture
In aggregation blood platelet separated with the blood platelet that do not assemble the second separation and concentration area, for detection of aggregation blood platelet quantity and
Second quantity detection zone of the quantity for the blood platelet that do not assemble.The blood platelet method for separating and concentrating can quickly, efficiently separate out
Blood platelet in blood sample.
The embodiment of the present invention provides a kind of reagent method of Platelet drug comprising following steps：
S1, it flows blood sample in the duct, dielectrophoresis force, the dielectric of application is applied to the blood sample sample in pipeline
The frequency of electrophoretic force is generally 500KHz-3MHz, separates blood platelet and other particles in blood sample.
S2, the quantity for detecting blood platelet, specific method are：Using Electrode with Electrochemical Impedance Spectroscopy, the electrochemistry resistance of blood platelet is detected
Anti- and frequency obtains blood platelet according to the corresponding relationship between the electrochemical impedance and frequency and platelet counts of blood platelet
Quantity.
S3, in blood platelet be added inducer stay for some time, assemble blood platelet sufficiently, obtain aggregation blood platelet and
The mixture of (not assembling) blood platelet.The inducer that the present embodiment uses includes collagen (COLL), adenosine diphosphate (ADP)
(ADP), one of adrenaline (EPN), arachidonic acid (ACA), ristomycin (RIST).
S4, flow the mixture of aggregation blood platelet and the blood platelet that do not assemble in the duct, to the mixture in pipeline
Apply dielectrophoresis force, the frequency of the dielectrophoresis force of application is generally 500KHz-3MHz, keeps the aggregation blood in mixture small
Plate is separated with the blood platelet that do not assemble.
The quantity of S5, the quantity of detection of aggregation blood platelet and the blood platelet that do not assemble, specific method are：It is hindered using electrochemistry
Anti- method detects blood platelet/aggregation blood platelet electrochemical impedance and frequency, is hindered according to blood platelet/aggregation blood platelet electrochemistry
The anti-and corresponding relationship between frequency and blood platelet/aggregation platelet counts, obtains blood platelet/aggregation blood platelet quantity.
S6, Platelet drug is added in the mixture of aggregation blood platelet and blood platelet, after obtaining drug effect
Assemble the mixture of blood platelet and blood platelet.Platelet drug existing related drugs on the market, are typically divided into four classes,
Respectively inhibit blood platelet arachidonic acid metabolic medicine (thromboxane A2 inhibitor：Aspirin), increase blood platelet inner ring nucleosides
Medicine (the Pimobendane of acid content：Dipyridmole (persantine)), specificity inhibit ADP activated blood platelet medicine, blood
Platelet fibrinogen deceptor (II b/ of GP, III a) antagonist.
S7, it flows the mixture after drug effect in the duct, dielectrophoresis force is applied to the mixture in pipeline, is applied
The frequency of the dielectrophoresis force added is generally 500KHz-3MHz, aggregation blood platelet and blood in the mixture after making drug effect
Platelet separates.
The quantity of aggregation blood platelet after S8, detection drug effect and the quantity of blood platelet, specific method are：Using electrification
Impedance method is learned, blood platelet/aggregation blood platelet electrochemical impedance and frequency are detected, according to blood platelet/aggregation blood platelet electrification
The corresponding relationship between impedance and frequency and blood platelet/aggregation platelet counts is learned, blood platelet/aggregation blood platelet number is obtained
Amount.According to the quantity variation of the quantity of aggregation blood platelet before and after dosing and blood platelet, that is, quickly detects blood platelet and drug is made
Assembling Behavior variation after, can calculate effect and effect of each drug to platelet aggregation.
The reagent method of the Platelet drug of the present embodiment is carried out in a pipeline, and pipeline is along its length
It successively include 8 regions, each region is corresponding to carry out an above-mentioned step, and so that blood sample or mixture is existed by air pump
Flowing in pipeline, i.e., from a next region of field flow orientation.
Shown in referring to figs. 2 and 3, the embodiment of the present invention also provides a kind of reagent based on above-mentioned Platelet drug
The reagent chip 100 of the Platelet drug of method comprising main body 110, main body 110 is interior to be equipped with three branches 120 and one
Pipeline 130, one end of three branches 120 are connected to the same end of pipeline 130 respectively, and three branches 120 are respectively used to pipe
Road 130 is passed through blood sample and sheath fluid, and usually intermediate branch 120 is passed through blood sample, and the branch 120 on both sides is all passed through
Sheath fluid.One end that pipeline 130 is connected with three branches 120 is divided into 8 regions to the other end, is followed successively by for making in blood sample
Blood platelet separated with other particles the first separation and concentration area 151, the first quantity detection zone for detecting platelet counts
152, the aggregation inducing area 153 for assembling blood platelet sufficiently, for making aggregation blood platelet in mixture and not assembling
The second separated separation and concentration area 171 of blood platelet, for the quantity of detection of aggregation blood platelet and the quantity for the blood platelet that do not assemble
The second quantity detection zone 172, reagent area 173, for making aggregation blood platelet and blood platelet in the mixture after drug effect
The quantity of separated third separation and concentration area 191 and quantity and blood platelet for detecting the aggregation blood platelet after drug effect
Third quantity detection zone 192.According to actual use situation, the diameter of pipeline 130 is 15-50um, the first separation and concentration area 151
Length be 500um-1mm, the length of the first quantity detection zone 152 is 500um-1mm, and the length in aggregation inducing area 153 is 1-
5mm, the length in the second separation and concentration area 171 are 500um-1mm, and the length of the second quantity detection zone 172 is 500um-1mm, examination
The length in medicine area 173 is 1-5mm, and the length in third separation and concentration area 191 is 500um-1mm, the length of third quantity detection zone 192
Degree is 500um-1mm.
In the present embodiment, the use being connected to respectively with three branches 120 far from one end of pipeline 130 is offered in main body 110
In injection port 141, the first reagent for reagent 1 (sheath fluid) to be added into pipeline 130 that blood sample is added into pipeline 130
Hole 142, the second reagent wells 143 for reagent 2 (sheath fluid) to be added into pipeline 130, are connected to aggregation inducing area 153, are used for
The inducer adding mouth 144 of inducer is added into pipeline 130, is connected to reagent area 173, for blood to be added into pipeline 130
The drug adding mouth 145 of platelet drugs with function, be connected to pipeline 130 far from one end of three branches 120, for taking out reagent after
Blood sample sample outlet 146.
In the present embodiment, pipeline 130 successively includes being detected by the first separation and concentration area 151, the first quantity along its length
Area 152 and aggregation inducing area 153 form the first segment pipe 150, the first connecting pipe 160, by the second separation and concentration area 171, the
The second segment pipe 170 that two quantity detection zones 172 and reagent area 173 form, the second connecting pipe 180, by third separation and concentration
The third segment pipe 190 that area 191 and third quantity detection zone 192 form, the first connecting pipe 160 and the second connecting pipe 180
On be separately provided for control pipeline 130 be opened and closed valve 161.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
The present embodiment provides a kind of reagent methods that the reagent chip 100 using Platelet drug carries out comprising
Following procedure：
S1, blood sample is added by injection port 141, reagent 1 is added by the first reagent wells 142, passes through the second reagent
Reagent 2 is added in hole 143, forms inhomogeneous field in the first separation and concentration area 151, by air pump make blood sample and reagent 1,
Reagent 2 flows respectively by the first separation and concentration area 151 being pooled in pipeline 130 along the length direction of pipeline 130, from
And dielectrophoresis force is applied to the blood sample sample in pipeline 130, the frequency of the dielectrophoresis force of application is generally 500KHz-
3MHz separates blood platelet and other particles in blood sample, flows at different rates, enters the first quantity detection zone
152。
S2, in the first quantity detection zone 152, using Electrode with Electrochemical Impedance Spectroscopy, detect the electrochemical impedance and frequency of blood platelet
Rate obtains the quantity of blood platelet, blood according to the corresponding relationship between the electrochemical impedance and frequency and platelet counts of blood platelet
Platelet enters aggregation inducing area 153.
S3, collagen is added in the blood platelet in aggregation inducing area 153 by inducer adding mouth 144 and stops one
The section time, assemble blood platelet sufficiently, obtain the mixture of aggregation blood platelet and blood platelet, opens in the first connecting pipe 130
Valve 161, so that mixture is entered the second separation and concentration area 171.
S4, inhomogeneous field is formed in the second separation and concentration area 171, makes the mixture for assembling blood platelet and blood platelet along pipe
The length direction in road 130 flows, so that dielectrophoresis force is applied to the mixture in pipeline 130, the dielectrophoresis force of application
Frequency is generally 500KHz-3MHz, separates aggregation blood platelet and blood platelet in mixture, flows at different rates, into
Enter to the second quantity detection zone 172.
S5, in the second quantity detection zone 172, using Electrode with Electrochemical Impedance Spectroscopy, detect blood platelet/aggregation blood platelet respectively
Electrochemical impedance and frequency, according to blood platelet/aggregation blood platelet electrochemical impedance and frequency and blood platelet/aggregation platelet count
Corresponding relationship between amount obtains blood platelet/aggregation blood platelet quantity, after detection, assembles blood platelet and blood platelet all
Mixture is formed again into reagent area 173.
S6, aggregation blood platelet and blood platelet by drug adding mouth 145 in reagent area 173 mixture in be added Ah
A woods is taken charge of, the mixture of aggregation blood platelet and blood platelet after obtaining aspirin effect is opened in the second connecting pipe 130
Valve 161, the mixture after acting on aspirin enter third separation and concentration area 191.
S7, inhomogeneous field is formed in third separation and concentration area 191, the mixture after acting on aspirin is along pipeline
130 length direction flowing, to apply dielectrophoresis force, the frequency of the dielectrophoresis force of application to the mixture in pipeline 130
Rate is generally 500KHz-3MHz, and aggregation blood platelet and the blood platelet in mixture after acting on aspirin separate, with difference
Speed flowing, enter third quantity detection zone 192.
S8, in third quantity detection zone 192, using Electrode with Electrochemical Impedance Spectroscopy, detect blood platelet/aggregation blood platelet respectively
Electrochemical impedance and frequency, according to blood platelet/aggregation blood platelet electrochemical impedance and frequency and blood platelet/aggregation platelet count
Corresponding relationship between amount obtains blood platelet/aggregation blood platelet quantity, takes out sample by outlet 146.Before dosing
Assemble the quantity of blood platelet and the quantity variation of blood platelet afterwards, effect and the function of aspirin on platelet aggregation can be calculated
Effect.
In conclusion the blood platelet method for separating and concentrating of the embodiment of the present invention can quickly, efficiently separate out blood sample
In blood platelet；The reagent method of the Platelet drug of the embodiment of the present invention can quickly detect blood platelet and drug effect
Assembling Behavior variation afterwards；The structure of the reagent chip of the Platelet drug of the embodiment of the present invention is simple, at low cost, can
Quickly carry out the reagent of Platelet drug.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of blood platelet method for separating and concentrating, which is characterized in that it includes the following steps：
It flows blood sample in the duct, dielectrophoresis force is applied to the blood sample sample in pipeline, makes the blood in blood sample
Platelet is separated with other particles, detects the quantity of blood platelet；
Inducer is added in the blood platelet, assembles blood platelet sufficiently, obtains the mixture of aggregation blood platelet and blood platelet；
Make to assemble blood platelet and the mixture of blood platelet flow in the duct, dielectrophoresis force is applied to the mixture in pipeline,
Separate aggregation blood platelet and blood platelet in mixture, the quantity of detection of aggregation blood platelet and the quantity of blood platelet.
2. blood platelet method for separating and concentrating according to claim 1, which is characterized in that detection blood platelet/aggregation blood platelet
The method of quantity be：Using Electrode with Electrochemical Impedance Spectroscopy, blood platelet/aggregation blood platelet electrochemical impedance and frequency are detected, according to
Corresponding relationship between blood platelet/aggregation blood platelet electrochemical impedance and frequency and blood platelet/aggregation platelet counts, obtains
Blood platelet/aggregation blood platelet quantity.
3. blood platelet method for separating and concentrating according to claim 1, which is characterized in that the frequency of the dielectrophoresis force of application
For 500KHz-3MHz.
4. blood platelet method for separating and concentrating according to claim 1, which is characterized in that the inducer includes collagen egg
One of white, adenosine diphosphate (ADP), adrenaline, arachidonic acid, ristomycin.
5. blood platelet method for separating and concentrating according to claim 1, which is characterized in that make blood sample by air pump or mix
Object is closed to flow in pipeline.
6. a kind of reagent method of Platelet drug, which is characterized in that it includes the following steps：
Using blood platelet method for separating and concentrating as described in claim 1, the quantity of aggregation blood platelet and the number of blood platelet are obtained
Amount；
Platelet drug is added in the mixture of aggregation blood platelet and blood platelet, the aggregation blood after obtaining drug effect is small
The mixture of plate and blood platelet；
Mixture after making drug effect flows in the duct, applies dielectrophoresis force to the mixture in pipeline, makees drug
Aggregation blood platelet and blood platelet in mixture after separate, and the quantity and blood of the aggregation blood platelet after detecting drug effect are small
The quantity of plate.
7. the reagent method of Platelet drug according to claim 6, which is characterized in that the Platelet medicine
Object is divided into four classes, respectively inhibition blood platelet arachidonic acid metabolic medicine, the medicine, special for increasing blood platelet cyclic nucleotide content
Property inhibit ADP activated blood platelet medicine, platelet fibrinogen receptor antagonist.
8. a kind of reagent core of the Platelet drug of the reagent method based on Platelet drug described in claim 1
Piece, which is characterized in that it includes main body, and three branches and a pipeline, one end of three branches are equipped in the main body
It is connected to respectively with the same end of the pipeline, one end that the pipeline is connected with three branches is followed successively by the other end for making blood
The first separation and concentration area that blood platelet in liquid sample is separated with other particles, the first quantity for detecting platelet counts are examined
Survey area, the aggregation inducing area for assembling blood platelet sufficiently, for separating aggregation blood platelet and blood platelet in mixture
The second separation and concentration area, the second quantity detection zone of quantity of quantity and blood platelet for detection of aggregation blood platelet, reagent
Area, the third separation and concentration area for separating the aggregation blood platelet in the mixture after drug effect with blood platelet and for examining
The quantity of aggregation blood platelet after surveying drug effect and the quantity third quantity detection zone of blood platelet.
9. the reagent chip of Platelet drug according to claim 8, which is characterized in that offered in the main body
The injection port that is connected to respectively with three branches far from one end of the pipeline, the first reagent wells, the second reagent wells, with the aggregation
Induce the inducer adding mouth of area's connection, the drug adding mouth being connected to the reagent area, with the pipeline far from three branches
One end connection outlet.
10. the reagent chip of Platelet drug according to claim 8, which is characterized in that the pipeline is along length
Direction successively includes being connected by the first separation and concentration area, the first quantity detection zone and aggregation inducing district's groups at the first segment pipe, first
Adapter tube road, by the second separation and concentration area, the second quantity detection zone and reagent district's groups at the second segment pipe, the second connecting pipe,
The third segment pipe being made of third separation and concentration area and third quantity detection zone, first connecting pipe and described second connect
The valve of control pipeline opening and closing is separately provided on adapter tube road.
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| CN110314713A (en) * | 2019-06-11 | 2019-10-11 | 南京理工大学 | A kind of biological molecule concentration device and its control method based on dielectrophoresis |
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